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Purification of hemoglobin by ion exchange chromatography in flow-through mode with PEG as an escort
Alternative TitleArtif. Cells Blood Substit. Immobil. Biotechnol.
Lu, XL; Zhao, DX; Su, ZG
2004
Source PublicationARTIFICIAL CELLS BLOOD SUBSTITUTES AND IMMOBILIZATION BIOTECHNOLOGY
ISSN1073-1199
Volume32Issue:2Pages:209-227
AbstractDevelopment of hemoglobin-based blood substitutes requires production of highly purified hemoglobin. Process of hemoglobin purification by ion exchange chromatography in flow-through mode was researched and optimized. Three kinds of media including, QMA Spherosil LS (Biosepra, France) and Q Sepharose Big Beads (Amersham Bioscience, Sweden), and an anion exchange membrane column, Mustang Q (PALL, USA) were investigated and compared. Adding polyethylene glycol (PEG) as an escort in ion exchange chromatography improved the purity and recovery, and the recovery in the chromatography was increased from 75 to 95%. The mechanism of PEG effects on chromatography was discussed. The optimal chromatography step, in combination with hypotonic dilution hemolyzing and membrane separation, formed an integrated hemoglobin purification process. The total recovery in the process was 87.6%. The activity of hemoglobin was well preserved: P-50 23.2 mmHg, and Hill coefficient 2.31. The product appeared as a single band in SDS-PAGE, and GF-HPLC showed only one peak. The purity of the prepared hemoglobin was more than 99.9%. The optimized process is time saving and suitable for large-scale preparation of hemoglobin to provide materials for further preparation of blood substitutes.; Development of hemoglobin-based blood substitutes requires production of highly purified hemoglobin. Process of hemoglobin purification by ion exchange chromatography in flow-through mode was researched and optimized. Three kinds of media including, QMA Spherosil LS (Biosepra, France) and Q Sepharose Big Beads (Amersham Bioscience, Sweden), and an anion exchange membrane column, Mustang Q (PALL, USA) were investigated and compared. Adding polyethylene glycol (PEG) as an escort in ion exchange chromatography improved the purity and recovery, and the recovery in the chromatography was increased from 75 to 95%. The mechanism of PEG effects on chromatography was discussed. The optimal chromatography step, in combination with hypotonic dilution hemolyzing and membrane separation, formed an integrated hemoglobin purification process. The total recovery in the process was 87.6%. The activity of hemoglobin was well preserved: P-50 23.2 mmHg, and Hill coefficient 2.31. The product appeared as a single band in SDS-PAGE, and GF-HPLC showed only one peak. The purity of the prepared hemoglobin was more than 99.9%. The optimized process is time saving and suitable for large-scale preparation of hemoglobin to provide materials for further preparation of blood substitutes.
KeywordHemoglobin Ion Exchange Chromatography Process Integration Protein Purification Blood Substitutes
SubtypeArticle
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine ; Technology
DOI10.1081/BIO-120037828
URL查看原文
Indexed BySCI
Language英语
WOS KeywordPOLYETHYLENE-GLYCOL ; SUBSTITUTES
WOS Research AreaBiotechnology & Applied Microbiology ; Engineering ; Materials Science
WOS SubjectBiotechnology & Applied Microbiology ; Engineering, Biomedical ; Materials Science, Biomaterials
WOS IDWOS:000222338200003
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Cited Times:18[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Version出版稿
Identifierhttp://ir.ipe.ac.cn/handle/122111/5004
Collection研究所(批量导入)
AffiliationChinese Acad Sci, Inst Proc Engn, Natl Key Lab Biochem Engn, Beijing 100080, Peoples R China
Recommended Citation
GB/T 7714
Lu, XL,Zhao, DX,Su, ZG. Purification of hemoglobin by ion exchange chromatography in flow-through mode with PEG as an escort[J]. ARTIFICIAL CELLS BLOOD SUBSTITUTES AND IMMOBILIZATION BIOTECHNOLOGY,2004,32(2):209-227.
APA Lu, XL,Zhao, DX,&Su, ZG.(2004).Purification of hemoglobin by ion exchange chromatography in flow-through mode with PEG as an escort.ARTIFICIAL CELLS BLOOD SUBSTITUTES AND IMMOBILIZATION BIOTECHNOLOGY,32(2),209-227.
MLA Lu, XL,et al."Purification of hemoglobin by ion exchange chromatography in flow-through mode with PEG as an escort".ARTIFICIAL CELLS BLOOD SUBSTITUTES AND IMMOBILIZATION BIOTECHNOLOGY 32.2(2004):209-227.
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